Journal: Cell reports

Article Title: The STRIPAK complex is required for radial sorting and laminin receptor expression in Schwann cells

doi: 10.1016/j.celrep.2025.115401

Figure Lengend Snippet:

Article Snippet: Membranes were then incubated with prey proteins GST-PAK-PBD, His-Strn3 (MyBioSource MBS1348331), or His-Mob4-StrepII (prepared from Addgene expression plasmid # 31592, http://n2t.net/addgene:31592 , RRID: Addgene_31592, a gift from Konrad Büssow) diluted to 100 nM in 5% BSA in TBS-T + 1 mM MgCl 2 overnight at 4°C [ ] .

Techniques: Generated, Virus, Recombinant, SYBR Green Assay, Protease Inhibitor, Plasmid Preparation, In Situ, BIA-KA, Mutagenesis, Software

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: TRAF3IP3 Induces ER Stress-Mediated Apoptosis with Protective Autophagy to Inhibit Lung Adenocarcinoma Proliferation.

doi: 10.1002/advs.202411020

Figure Lengend Snippet: Figure 7. TRAF3IP3 interacts with STRN3 and promotes STRN3 localization of endoplasmic reticulum. A) STRN3 was identified as the binding partner of TRAF3IP3 by two replicate co-IP mass spectrometry experiments. B) Molecular docking of TRAF3IP3 and STRN3. The purple molecule stands for TRAF3IP3, and the sky-blue molecule stands for STRN3. C) Exogenous reciprocal coimmunoprecipitation (co-IP) of TRAF3IP3 and STRN3 in 293T cells overexpressing Flag-TRAF3IP3 and/or HA-STRN3. D) Immunofluorescence staining of DAPI (blue), TRAF3IP3 (red) and STRN3 (green) in LUAD cell lines. E) The colocalization curves showed the colocalization level of STRN3 and TRAF3IP3 in the representative image. F) Schematic representation of wild-type TRAF3IP3 and TRAF3IP3 deletion truncations. G Lysates from HEK293T cells transfected with HA-tagged STRN3 and Flag-tagged wild-type TRAF3IP3 or TRAF3IP3 deletion truncations (1-235, 236–450, 451–520, 521–551) were collected for coimmunoprecipitation and immunoblotting. H) A549 cell was transfected with indicated expression vectors, and STRN3, GRP78, ATF4, and Bax levels were detected by western blotting. I) Wild-type TRAF3IP3 or deletion truncations were transfected into A549 cells, and confocal microscopy analysis of GRP78, STRN3 and DAPI was conducted. J) The colocalization curves showed the colocalization level of GRP78 and STRN3 in the representative image. K,L) A549 cell was transfected with control vector, wild-type TRAF3IP3, specific containing transmembrane domain, and transmembrane domain-deleted (rTM) plasmids; cell lysates were subjected to immunoblot with indicated antibodies.

Article Snippet: Thereafter, the cells were permeabilized in 0.2% Triton X100 (Sigma-Aldrich, China) at 4 °C for 10 min and blocked in PBSB (4% BSA) for 1 h. The cells were then fluorescently double-stained with primary antibodies against TRAF3IP3(Santa, sc-398895), STRN3 (Abclonal, A6756) or GRP78(Proteintech, 66574-1-lg) at 4 °C overnight.

Techniques: Binding Assay, Co-Immunoprecipitation Assay, Mass Spectrometry, Staining, Transfection, Western Blot, Expressing, Confocal Microscopy, Control, Plasmid Preparation

Journal: The Journal of Biological Chemistry

Article Title: Reactivating Hippo by drug compounds to suppress gastric cancer and enhance chemotherapy sensitivity

doi: 10.1016/j.jbc.2024.107311

Figure Lengend Snippet: Identification of DSF as an inhibitor of the PP2Aa–STRN3 interaction. A , illustration of the first-round AlphaScreen to find out compounds disrupting PP2Aa–STRN3 interaction. B , workflow of four step-by-step compound screening and confirmation strategies. N represents the number of compounds after each screening step. C , result of inhibition rate for disrupting PP2Aa–STRN3 interaction in the first round AlphaScreen. Dash line refers to inhibition efficiency higher than 60% (n = 47 for red circle ). D , result of the second round AlphaScreen to kick out false positive hits which disrupted interactions between protein and donor/acceptor beads instead of breaking the PP2Aa–STRN3 protein–protein interaction. Vertical dash line refers to inhibition efficiency higher than 50% while horizontal dash line refers to false positive probability less than 35% (n = 9). E , IC 50 values of nine candidate compounds in HGC-27 cells. The IC 50 values are less than 5 μM highlighted in red . F , GST pulldown assay to confirm the disruption effect of PP2Aa–STRN3 association with the indicated 10 μM nine compounds treatment. STRN3 refers to amino acid 64 to 145, hereafter. G , co-IP analysis of the interaction between HA-PP2Aa and Flag-STRN3 in HEK293A cells treated with indicated dose of DSF. Co-IP, co-immunoprecipitation; DSF, disulfiram; HS, His-Sumo tag; ev, empty vector.

Article Snippet: To identify drug compounds which can mimic the SHAP peptide–induced Hippo reactivation strategy via disrupting the PP2Aa–STRN3 interaction ( A , previous work) , we applied a protein–protein interaction-based AlphaScreen platform to screen an FDA-approved drug library containing 3095 chemicals (TargetMol inhibitors) ( A ).

Techniques: Amplified Luminescent Proximity Homogenous Assay, Inhibition, GST Pulldown Assay, Disruption, Co-Immunoprecipitation Assay, Immunoprecipitation, Plasmid Preparation

Journal: The Journal of Biological Chemistry

Article Title: Reactivating Hippo by drug compounds to suppress gastric cancer and enhance chemotherapy sensitivity

doi: 10.1016/j.jbc.2024.107311

Figure Lengend Snippet: DSF competes with STRN3 for PP2Aa binding. A , detection the DSF–PP2Aa binding affinity and its inhibitory effect on PP2Aa–STRN3 association by MST assay. B , virtual docking of DSF binding to first HEAT repeat of the PP2Aa. The PDB ID for crystal structure of the PP2Aa–STRN3 complex is 6IUR. PP2Aa is colored in green and shown as cartoon. DSF is shown as sticks and colored in red . The right panel illustrates the binding site of DSF to PP2Aa. C , illustration of the occupied position of DSF in PP2Aa–STRN3 complex. D , surface structure to highlight the binding pocket (V15, A41) of DSF towards PP2Aa. PP2Aa is shown as surface. E , MST assay to show the incapability of PP2Aa mutants in binding to DSF. PP2A V15/A41D : PP2Aa mutant variant with V15 and A41 substituted with a glutamate. F , model for DSF competing with STRN3 for PP2Aa binding. DSF, disulfiram; MST, microscale thermophoresis.

Article Snippet: To identify drug compounds which can mimic the SHAP peptide–induced Hippo reactivation strategy via disrupting the PP2Aa–STRN3 interaction ( A , previous work) , we applied a protein–protein interaction-based AlphaScreen platform to screen an FDA-approved drug library containing 3095 chemicals (TargetMol inhibitors) ( A ).

Techniques: Binding Assay, Mutagenesis, Variant Assay, Microscale Thermophoresis

Journal: The Journal of Biological Chemistry

Article Title: Reactivating Hippo by drug compounds to suppress gastric cancer and enhance chemotherapy sensitivity

doi: 10.1016/j.jbc.2024.107311

Figure Lengend Snippet: TH acts as a structural analog of DSF to inhibit GC. A , the chemical structures of DSF and TH. B , GST pulldown assay to demonstrate the dose-dependent disruption effect of TH on PP2Aa–STRN3 association. The dosage for TH is 0, 2.5, 5, 10 μM, respectively. C , MST assay to show the interruption effect and association between TH and PP2Aa. D , virtual docking of TH binding to first HEAT repeat of the PP2Aa-occupied STRN3-binding site. STRN3 was colored by cyan while PP2Aa was colored by green . E , surface structure to highlight the binding pocket (V15, A41) of TH towards PP2Aa. PP2Aa was shown as surface. F , detection of the disruption effect of PP2Aa mutant in binding with DSF by MST assay. PP2A Mut : PP2Aa mutant variant with V15 and A41 substituted with a glutamate. G , in vitro dephosphorylation assay to confirm dose-dependently effects of TH on the phosphatase activity of PP2Aa/c core enzyme using pMST2 (T180) as a substrate. H , GSEA analysis of YAP signature genes in TH-treated HGC-27 cells for 48 h. Normalized enrichment score (NES) and FDR values are indicated. I , immunoblots of the phosphorylation level of Hippo components in HGC-27 cells treated with or without TH (200 nM). J , relative mRNA levels of CTGF in GES-1 and HGC-27 cells treated with TH for 48 h (n = 3). Significance was tested using one-way ANOVA, followed by the Tukey’s post hoc test. ∗∗ p < 0.01; ∗∗∗ p < 0.001. K , cell viability of two different cell lines (HGC-27 and GES-1) with TH treatment (n = 3). L , cell viability of HGC-27 cells treated with DSF or TH (n = 3). M , cell viability of HGC-27 cells treated with DSF/TH or DSF/TH-Cu complex (n = 3). Data are presented as means ± SD. The data were analyzed using one-way ANOVA, followed by the Tukey’s post hoc test. ∗∗ p < 0.01; ∗∗∗ p < 0.001. N , the colony formation of HGC-27 cells treated with TH of different doses (n = 3). The right bar figure represents the statistics of colony numbers. Data are presented as means ± SD. The data were analyzed using one-way ANOVA, followed by the Tukey’s post hoc test. ∗∗ p < 0.01; ∗∗∗∗ p < 0.0001. O , killing efficiencies of AGS cells treated with 0.1 μM TH and 10 μM cisplatin alone or in combination. Significance was tested using one-way ANOVA, followed by the Tukey’s post hoc test. ∗∗ p < 0.01; ∗∗∗ p < 0.001; n.s., no significance. DSF, disulfiram; GC, gastric cancer; GSEA, gene set enrichment analysis; MST, microscale thermophoresis.

Article Snippet: To identify drug compounds which can mimic the SHAP peptide–induced Hippo reactivation strategy via disrupting the PP2Aa–STRN3 interaction ( A , previous work) , we applied a protein–protein interaction-based AlphaScreen platform to screen an FDA-approved drug library containing 3095 chemicals (TargetMol inhibitors) ( A ).

Techniques: GST Pulldown Assay, Disruption, Binding Assay, Mutagenesis, Variant Assay, In Vitro, De-Phosphorylation Assay, Activity Assay, Western Blot, Phospho-proteomics, Microscale Thermophoresis

Journal: The Journal of Biological Chemistry

Article Title: Reactivating Hippo by drug compounds to suppress gastric cancer and enhance chemotherapy sensitivity

doi: 10.1016/j.jbc.2024.107311

Figure Lengend Snippet: A schematic illustration of the rationale of Hippo restoration for GC treatment. The STRN3 acts as a bridge to link PP2A phosphatase and MST1/2 kinases, mediating the dephosphorylation of MST1/2. As such, the activity of MST1/2 is properly controlled. In tumors, STRN3 upregulation leads to enhanced inhibition of MST1/2 (Hippo off), resulting in the defective of LATS1/2-mediaed YAP dephosphorylation, transactivation of TEAD-dependent gene transcription, and GC development. Meanwhile, such sequestration of MST1/2 in the cytoplasm leads to elevated DNA repair capacity and drug resistance in response to chemotherapy ( left panel ). The FDA-approved drugs such as DSF and its structural analog TH, as well as CX, were concluded from the current work enabling to either block the PP2Aa–STRN3 or STRN3–MST1/2 interaction, respectively, and therefore restore the activity of MST1/2 (Hippo restoration) and in this way serve as a promising therapy for GC ( right panel ). CX, CX-6258; DSF, disulfiram; GC, gastric cancer.

Article Snippet: To identify drug compounds which can mimic the SHAP peptide–induced Hippo reactivation strategy via disrupting the PP2Aa–STRN3 interaction ( A , previous work) , we applied a protein–protein interaction-based AlphaScreen platform to screen an FDA-approved drug library containing 3095 chemicals (TargetMol inhibitors) ( A ).

Techniques: De-Phosphorylation Assay, Activity Assay, Inhibition, Blocking Assay

Journal: Chinese Journal of Cancer Research

Article Title: Striatins and STRIPAK complex partners in clinical outcomes of patients with breast cancer and responses to drug treatment

doi: 10.21147/j.issn.1000-9604.2023.04.04

Figure Lengend Snippet: PCR primers

Article Snippet: Lentiviral short hairpin RNA (shRNA) targeting human STRN (SC-37649v) and STRN3 (SG2NA) (SC-37647v) and small interfering (siRNA) targeting STRN (SC-37649) and STRN3 (SG2NA) (SC-37647) were purchased from Santa Cruz Biotechnologies Inc. Each knockdown included three sets of siRNAs.

Techniques:

Journal: Chinese Journal of Cancer Research

Article Title: Striatins and STRIPAK complex partners in clinical outcomes of patients with breast cancer and responses to drug treatment

doi: 10.21147/j.issn.1000-9604.2023.04.04

Figure Lengend Snippet: STRIPAK and patients’ OS and DFS, Cardiff data by Cox regression

Article Snippet: Lentiviral short hairpin RNA (shRNA) targeting human STRN (SC-37649v) and STRN3 (SG2NA) (SC-37647v) and small interfering (siRNA) targeting STRN (SC-37649) and STRN3 (SG2NA) (SC-37647) were purchased from Santa Cruz Biotechnologies Inc. Each knockdown included three sets of siRNAs.

Techniques:

Journal: Chinese Journal of Cancer Research

Article Title: Striatins and STRIPAK complex partners in clinical outcomes of patients with breast cancer and responses to drug treatment

doi: 10.21147/j.issn.1000-9604.2023.04.04

Figure Lengend Snippet: Patient’s drug response, tumor pathological responses to chemotherapies *

Article Snippet: Lentiviral short hairpin RNA (shRNA) targeting human STRN (SC-37649v) and STRN3 (SG2NA) (SC-37647v) and small interfering (siRNA) targeting STRN (SC-37649) and STRN3 (SG2NA) (SC-37647) were purchased from Santa Cruz Biotechnologies Inc. Each knockdown included three sets of siRNAs.

Techniques:

Journal: Chinese Journal of Cancer Research

Article Title: Striatins and STRIPAK complex partners in clinical outcomes of patients with breast cancer and responses to drug treatment

doi: 10.21147/j.issn.1000-9604.2023.04.04

Figure Lengend Snippet: Patient’s drug response, 5-year RFS response to chemotherapies *

Article Snippet: Lentiviral short hairpin RNA (shRNA) targeting human STRN (SC-37649v) and STRN3 (SG2NA) (SC-37647v) and small interfering (siRNA) targeting STRN (SC-37649) and STRN3 (SG2NA) (SC-37647) were purchased from Santa Cruz Biotechnologies Inc. Each knockdown included three sets of siRNAs.

Techniques:

Journal: Chinese Journal of Cancer Research

Article Title: Striatins and STRIPAK complex partners in clinical outcomes of patients with breast cancer and responses to drug treatment

doi: 10.21147/j.issn.1000-9604.2023.04.04

Figure Lengend Snippet: STRN3 transcript expression level and patients’ response to chemotherapies in subtypes of breast cancer ( * by Mann-Whitney U test)

Article Snippet: Lentiviral short hairpin RNA (shRNA) targeting human STRN (SC-37649v) and STRN3 (SG2NA) (SC-37647v) and small interfering (siRNA) targeting STRN (SC-37649) and STRN3 (SG2NA) (SC-37647) were purchased from Santa Cruz Biotechnologies Inc. Each knockdown included three sets of siRNAs.

Techniques: Expressing

Journal: Chinese Journal of Cancer Research

Article Title: Striatins and STRIPAK complex partners in clinical outcomes of patients with breast cancer and responses to drug treatment

doi: 10.21147/j.issn.1000-9604.2023.04.04

Figure Lengend Snippet: Expression of STRN and STRN3 in breast cancer cell lines. (A) qPCR confirmation of KDs. Semi-quantitative analysis of the relative gene expression of STRN and STRN3 in the four breast cancer cell lines, MDA-MB-231, MDA-MB-361, SKBR3, MCF-7. The fold change was acquired by 2−∆∆Ct which is a formula used to calculate the relative fold change expression when performing qPCR. Unpaired t-test was performed to statistically analyze the degree of KDs. *, P<0.05; **, P<0.01; ***, P<0.001; ****, P<0.0001; (B) Western blot shows the STRN and STRN3’s protein expression, respectively in WT(+) and KD(−) MDA-MB-231, MDA-MB-361 and SKBR3 cell lines. The corresponded protein expression of the housekeeping gene, GAPDH, in each cell model is also demonstrated. qPCR, quantitative real-time polymerase chain reaction; WT, wild type; KD, knockdown.

Article Snippet: Lentiviral short hairpin RNA (shRNA) targeting human STRN (SC-37649v) and STRN3 (SG2NA) (SC-37647v) and small interfering (siRNA) targeting STRN (SC-37649) and STRN3 (SG2NA) (SC-37647) were purchased from Santa Cruz Biotechnologies Inc. Each knockdown included three sets of siRNAs.

Techniques: Expressing, Gene Expression, Western Blot, Real-time Polymerase Chain Reaction, Knockdown

Journal: Chinese Journal of Cancer Research

Article Title: Striatins and STRIPAK complex partners in clinical outcomes of patients with breast cancer and responses to drug treatment

doi: 10.21147/j.issn.1000-9604.2023.04.04

Figure Lengend Snippet: Cell adhesiveness and migration of MDA-MB-231 cells with different STRNs expressions, namely MDA-MB-231 WT, MDA-MB-231 STRN-KD and MDA-MB-231 STRN3-KD. (A−D) adhesion assay; (E−H) migration assay. From left to right, (A,E) 2D graph compares cell responses among cell models detected at 4,000 Hz (resistance with standard errors); (B,F) 3D graph of cell responses for WT cells; (C,G) 3D graph of cell responses for STRN-KD cells; (D,H) 3D graph of cell responses for STRN3-KD cells. The 3D figures indicate cell responses (Z-axis, normalized resistance in ohms) over time (X-axis, hours) and across multiple frequencies (Y-axis, Hz). All resistance detected was normalized with the initial result acquired. WT, wild type; KD, knockdown.

Article Snippet: Lentiviral short hairpin RNA (shRNA) targeting human STRN (SC-37649v) and STRN3 (SG2NA) (SC-37647v) and small interfering (siRNA) targeting STRN (SC-37649) and STRN3 (SG2NA) (SC-37647) were purchased from Santa Cruz Biotechnologies Inc. Each knockdown included three sets of siRNAs.

Techniques: Migration, Cell Adhesion Assay, Knockdown

Journal: Chinese Journal of Cancer Research

Article Title: Striatins and STRIPAK complex partners in clinical outcomes of patients with breast cancer and responses to drug treatment

doi: 10.21147/j.issn.1000-9604.2023.04.04

Figure Lengend Snippet: Cell adhesiveness and migration of MDA-MB-361 cells with different STRNs expressions, namely MDA-MB-361 WT, MDA-MB-361 STRN-KD and MDA-MB-361 STRN3-KD. (A−D) adhesion assay; (E−H) migration assay. From left to right, (A,E) 2D graph compares cell responses among cell models detected at 4,000 Hz (resistance with standard errors); (B,F) 3D graph of cell responses for WT cells; (C,G) 3D graph of cell responses for STRN-KD cells; (D,H) 3D graph of cell responses for STRN3-KD cells. The 3D figures indicate cell responses (Z-axis, normalized resistance in ohms) over time (X-axis, hours) and across multiple frequencies (Y-axis, Hz). All resistance detected was normalized with the initial result acquired. WT, wild type; KD, knockdown.

Article Snippet: Lentiviral short hairpin RNA (shRNA) targeting human STRN (SC-37649v) and STRN3 (SG2NA) (SC-37647v) and small interfering (siRNA) targeting STRN (SC-37649) and STRN3 (SG2NA) (SC-37647) were purchased from Santa Cruz Biotechnologies Inc. Each knockdown included three sets of siRNAs.

Techniques: Migration, Cell Adhesion Assay, Knockdown

Journal: Chinese Journal of Cancer Research

Article Title: Striatins and STRIPAK complex partners in clinical outcomes of patients with breast cancer and responses to drug treatment

doi: 10.21147/j.issn.1000-9604.2023.04.04

Figure Lengend Snippet: Cell adhesiveness and migration of MCF-7 cells with different STRNs expressions, namely MCF-7 WT, MCF-7 STRN-KD and MCF-7 STRN3-KD. (A−D) adhesion assay; (E−H) migration assay. From left to right, (A,E) 2D graph compares cell responses among cell models detected at 4,000 Hz (resistance with standard errors); (B,F) 3D graph of cell responses for WT cells; (C,G) 3D graph of cell responses for STRN-KD cells; (D,H) 3D graph of cell responses for STRN3-KD cells. The 3D figures indicate cell responses (Z-axis, normalized resistance in ohms) over time (X-axis, hours) and across multiple frequencies (Y-axis, Hz). All resistance detected was normalized with the initial result acquired. WT, wild type; KD, knockdown.

Article Snippet: Lentiviral short hairpin RNA (shRNA) targeting human STRN (SC-37649v) and STRN3 (SG2NA) (SC-37647v) and small interfering (siRNA) targeting STRN (SC-37649) and STRN3 (SG2NA) (SC-37647) were purchased from Santa Cruz Biotechnologies Inc. Each knockdown included three sets of siRNAs.

Techniques: Migration, Cell Adhesion Assay, Knockdown

Journal: Chinese Journal of Cancer Research

Article Title: Striatins and STRIPAK complex partners in clinical outcomes of patients with breast cancer and responses to drug treatment

doi: 10.21147/j.issn.1000-9604.2023.04.04

Figure Lengend Snippet: Cell adhesiveness and migration of SKBR3 cells with different STRNs expressions, namely SKBR3 WT, SKBR3 STRN-KD and SKBR3 STRN3-KD. (A−D) adhesion assay; (E−H) migration assay. From left to right, (A,E) 2D graph compares cell responses among cell models detected at 4,000 Hz (resistance with standard errors); (B,F) 3D graph of cell responses for WT cells; (C,G) 3D graph of cell responses for STRN-KD cells; (D,H) 3D graph of cell responses for STRN3-KD cells. The 3D figures indicate cells responses (Z-axis, normalized resistance in ohms) over time (X-axis, hours) and across multiple frequencies (Y-axis, Hz). All resistance detected was normalized with the initial result acquired. WT, wild type; KD, knockdown.

Article Snippet: Lentiviral short hairpin RNA (shRNA) targeting human STRN (SC-37649v) and STRN3 (SG2NA) (SC-37647v) and small interfering (siRNA) targeting STRN (SC-37649) and STRN3 (SG2NA) (SC-37647) were purchased from Santa Cruz Biotechnologies Inc. Each knockdown included three sets of siRNAs.

Techniques: Migration, Cell Adhesion Assay, Knockdown

Journal: Chinese Journal of Cancer Research

Article Title: Striatins and STRIPAK complex partners in clinical outcomes of patients with breast cancer and responses to drug treatment

doi: 10.21147/j.issn.1000-9604.2023.04.04

Figure Lengend Snippet: IC 50 of chemo-drugs tests

Article Snippet: Lentiviral short hairpin RNA (shRNA) targeting human STRN (SC-37649v) and STRN3 (SG2NA) (SC-37647v) and small interfering (siRNA) targeting STRN (SC-37649) and STRN3 (SG2NA) (SC-37647) were purchased from Santa Cruz Biotechnologies Inc. Each knockdown included three sets of siRNAs.

Techniques: